This proposal presents a five-year research career development program focused on the study of HIV-1 eradication at the interface between virology and immunology. The candidate is currently an Instructor of Medicine at Harvard Medical School in the Division of Infectious Diseases at the Brigham and Women's Hospital. The outlined proposal builds on the candidate's previous research and clinical experience and integrates the two distinct domains of expertise represented by her mentor team of Dr. Galit Alter and Dr. Daniel Kuritzkes. The proposed experiments and didactic work will position the candidate with a unique set of cross-disciplinary skills that will enable her transition to independence as a physician scientist working in the field of HIV eradication. Despite remarkable advances in the medical management of HIV-1 infection, the epidemic continues to present clinical, social and economic challenges on a global scale. Strategies to interrupt and contain the epidemic are urgently needed. The recent description of isolated cases of functional cure of HIV-1 infection has turned attention to the possibility of virus eradication to achieve thi goal. The feasibility of this approach depends on both the development of targeted agents for HIV-1 latency reversal and on the successful elimination of cells harboring reactivating virus. This proposal focuses on the latter question, with the goal of developing a novel approach to enhance immune clearance of the HIV-1 reservoir. In place of the traditional focus on epitope or antigen specific adaptive immune responses, this proposal seeks to use the innate immune natural killer (NK) cell population to rapidly identify generic signals of stress and infection on reservoir cells through the NKG2D receptor. The studies evaluate both the direct effects of HIV-1 transcription and the influence of exposure to latency reversal agents on the expression of cellular stress signals and the functional competence of NK cells. This dual approach balances a focus on reactivation with an eye to preserving immune function. Further, to enhance NK mediated detection of reservoir cells the studies proposed will also test the efficacy of blocking the HIV-1 protein Nef to abrogate its immune evasion functions. Specific aims of this proposal include 1) Define the expression of stress molecules induced by latency reversal agents (LRAs) on HIV-1 reservoir cells 2) Determine the consequences of LRA exposure for NK cell phenotype and function 3) Enhance NK cell function by disabling viral immune evasion, leading to reduction of the HIV-1 reservoir. Positive results from these studies would provide the foundation for early clinical trials of a combined strategy of latency reversal and Nef blockade.